Centrifugal separation techniques for serum lipoproteins.
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Last Update: 2020-10-22
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Source: Internet
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Author: User
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serum
lipoprotein is an uneassuming complex consisting of lipids in the blood and certain specific
proteins
in the blood. Because of its different ratio of fat to lipoprotein, the density range is 0.96g/ml or less to 1.21g/ml. Paper electrophoresis shows four bands, namely: milky particles, very low density lipoprotein, LDL, HDL. Each lipoprotein can also be divided into more sub-groups.
electrophoresis method is more successful in separating high lipoprotein-containing groups (HDL, VHDL), while centrifugal separation methods are suitable for serum lipid
protein separation
.
, gradient materials:
serum lipoprotein density is low, commonly used gradient materials are Nacl, NaI, Kbr, KI, etc., commonly used buffer for ED.
II, pre-separation:
1, blood cell and serum separation:
centrifugal parameters: whole blood, angle turn, 3,000rpm×20min
Results: precipitation as blood cells, upper serum.
2, celiac grain separation: fasting for more than 12 hours in human plasma milk grain content is very small, has been eaters because of its plasma contains high celiac grain, should first centrifuge to remove celiac granules.
centrifugal parameters: 4 ×10 (g×min), 10 degrees C. All turn speeds are available.
gradient fluid configuration:
centrifuge tube
lower 3/4 volume plus plasma, upper 1/4 volume plus 0
5MnaCl plus 0.3M
EDTA
,ph7.4
result: milky granules floating, suction.
3, serum protein separation: remove globulin, albumin and other proteins.
centrifuge parameters: (2. 5-3) ×10 (g× hr), all kinds of turn heads are available. e.g. corner turn 70,000rpm× 6hrs, 10C
configuration: tube capacity 1/3 serum, 2/3 1. 31g/ml, NaCl plus NaBr, stirred with a final density of 1. 21g/ml
result: 1/6 volume of upper tube is serum lipoprotein, lower 5/6 is other proteins.
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